SUBSTANCE: blood plasma is examined in 4 minutes after the beginning of spontaneous red blood cell aggregation for free red blood cell count and cell count in aggregates. A percentage of non-aggregated red blood cells (PNA RBC) by formula PNA RBC=FRBSC×100/(TRBCA+FRBSC) wherein FRBSC is the free red blood cell count, TRBCA are total red blood cells in aggregates. If the PNA RBC is 56 to 30%, I degree of severity is stated, 30% to 4% - II degree of severity, less than 4% - III degree of severity.

EFFECT: use of the invention enables objectifying and increasing precision of evaluation of red blood cell aggregation, evaluating an intensity of patient's microcirculation disorders in a relatively short time, and thereby ensuring well-timed adequate complex of therapeutic measures or corrected therapy.

3 ex

The invention relates to medicine, namely to internal medicine, and can be used for rapid assessment of disorders of microcirculation.

In the last decade, both abroad and in Russia, significant progress has been made in the creation of fundamentally new approaches in the study of rheological properties of blood in clinical practice. The need for simple and informative diagnostic tests for detection of intravascular disorders of microcirculation is determined by the needs of practical health care in the problem solving methodology of diagnosis and correction of hemorheological disorders in patients with different pathologies, as well as to predict outcomes and monitoring the effectiveness of therapy.

One of the most important pathological phenomena that occur in the microcirculation system, is an intravascular aggregation of blood cells /mainly erythrocytes/(Firsov N.N., Lassie IV, Priezzhev A.V. Modern problems of whole blood aggregometry. Thrombosis, hemostasis and rheology. 2000, №2 (2), P.9-11; Ants L.V., I.A. Tikhomirov, D.V. Borisov analysis of the impact of plasma and cellular factors on the aggregation of erythrocytes of different age populations. Human physiology, 2002, V.28, No. 4, S-148). In this regard, the current study agregats the organizational capacity of erythrocytes and its correct interpretation.

The basis of the pathogenesis of many diseases and their complications are the processes violations of aggregation of red blood cells, initiating the development of thrombosis in the vessels. The need to study aggregation of erythrocytes is apparent and standardized instruments are missing. In different laboratories, there are various ways of assessing the aggregation of red blood cells, which complicates the interpretation of the resulting data. Frequently used method for quantitative determination of RBC aggregation is a photometric method for detecting decrease in the intensity of light scattered by blood after cessation of stirring the sample in a special cuvettes. Devices using this effect have been developed in our country (Levtov VA, Levkovich YU.N., Ashkenazi IA, I. Potapova About the study of the aggregation properties of blood. - Human physiology, 1978, V. 4, No. 3, s-513.; Lysov VA, Belousov SHE, Parfenov, AS the Method of determining the aggregation of erythrocytes. - Cardiology, 1978, No. 8, p.15-21; Sageline, GI Nazarenko, V. Ssize. Clinical aspects of the microcirculation. L., Medicine, 1985, s-106)and abroad (Schmid-Schnnbein N., Eline £A, Tolger E, Velocity of red cell aggregation (RCA): photometric determination of the half-teme and aggregation constant.-Bibl. Anat, 1975. N 13, p.71-92; Firsov N. N., A. V. Priezzhev, Ryaboshapka O., Sirko J.V. Aggregation properties of erythrocytes of whole blood under shear stress by backscattering nephelometry. Proc.of SPIE, 1993, v. 1884, p.283-290). However, dannysoar requires special equipment and involves complex calculations,
so not allows the physician to quickly assess the severity of disturbances of microcirculation of the patient and, accordingly, to make adjustments in treatment. The lack of standardized instruments leads to differences in the geometry of the flow and in the ways of the Desk leaving a blood sample light that leads to the difference of the obtained results.

Widely known direct optical method for the determination of RBC aggregation, recommended for use by who expert Committee for standardization in Hematology (International committee for standardization in hematology (expert panel on blood theology). Guidelines on selection of laboratory tests for monitoring the acute phase response. J. Clin. Pathol. 1988., V.41, P.1203-1212). This method is convenient and simple in execution and meets the following requirements: erythrocyte aggregation is taken into account in the conditions excluding its artificial amplification, the aggregation is taken into account when the standard initial volumetric ratio of the mass of red blood cells and plasma, approaching physiological, for cultivation plasma-suspended red blood cells (and at the same time "fixing" units) use the same suspensorium environment (plasma±aggregation substance), quantitative determination of aggregation in the cell (Goriaev) allows you to simultaneously estimate the size and shape of aggregates visually (Fernandez de CM. et al., 1989).

To improve the measurement accuracy of the detail of measured parameters, evaluation vergennes the violations and the aggregation of erythrocytes have a question about the classification of the aggregation violations by severity.
Attempt of classification of hemorheological disorders described in Firsova (Firsov N.N., Korotayev T.V., Vislova M.A. classification of the severity of hemorheological devices. Sat. "Rheological studies in medicine" (v.2), M, 2000, s-141). Taking as a prototype of the proposed Firsova method of separation of hemorheological disorders in degree I, II and III severity, we offer as a defining indicator of the degree of aggregation of violations, take the percentage of non-aggregated RBCs. This indicator is available and easy to identify in any laboratory, quite informative and does not require high material costs.

In the work of N.N. Firsova (2000) describes the principle of separation of severity. The basis of the average value (M), obtained from healthy donors, and standard deviation (σ), which determines the scattering parameter around the mean. Standard deviation is a parameter that reliably separates one average value from the other, a distance doubled its size (2σ). In a normal distribution in the interval M±σ is about 70% of all cases, therefore, the overlap between neighboring distributions is only 15%.

The technical result of the proposed method of assessing the severity of the violation aggregation Erythro itow is
that as a defining indicator of the severity of the aggregation disorders, calculate the percentage of non-aggregated RBCs (POE) according to the formula, and when POE from 56 to 30% I set severity, from 30% to 4% - II severity, less than 4% - III severity (see table).

A method of evaluating the severity of the violation aggregation of erythrocytes is that stable blood after 15 minutes of centrifugation at 3000 rpm divided by the plasma and the formed elements. The introduction of 0.2 ml were excavated plasma in plastic tablets, then add 0,02 ml washed in phosphate buffer RBC mass. After 4 minutes after the onset of spontaneous aggregation was carried out by counting the number of free (non-aggregated) of red blood cells using a light microscope in the cell Goryaeva, as well as the number of cells in aggregates. Counting was performed in 2 large (32 small) squares camera Goryaeva. Determined the percentage of non-aggregated RBCs (POE) according to the formula POE=CTU×100/(sea+CTU), where CTU - free erythrocytes, sea - sum of all cells in the aggregates.

Using the inventive method us when hemorheological survey 66 healthy people identified the average value and the standard deviation of the percentage of non-aggregated RBCs, sostaviv what I 69,0±13,0%.
Thus, the distribution of the severity of the following: rate of 82%-56%, I degree 56%-30%, II degree 30%-4%, third degree less than 4%. Among surveyed 66 healthy individuals POE is within the normal range were observed in 42 people (64%), I severity - 24 (36%).

Of the 86 patients with essential hypertension without organs targets normal POE was observed in 24 (28%>), I the degree of violation of aggregation of erythrocytes was observed in 49 (57%), II degree-13 (15%). Of the 75 patients with ischemic heart disease and myocardial infarction normal POE was observed in 6 patients (8%), first degree violation of aggregation of erythrocytes was observed in 37 people (49%), II degree-30 people (40%), grade III in 2 (3%)

Clinical example 1

Patient L., 43 years old were treated in the medical ward of the clinic Yugma GOU VPO University with 20.10.2007, 4.11.2007, with a diagnosis of hypertension Art. I, AG 2 tbsp., dyslipidemia, risk 3. Hypertensive history 2 years.

When hemorheological examination level POE was 42,86%, which refers to I the severity of the aggregation disorders. Other rheological parameters are as follows: average unit size - 6,07; the rate of aggregation is 1.91; blood viscosity at the shear rate of 200 s-1- 4.5 MPa·s, at a speed of 50-1-5,5 MPa·s at a shear rate of 10-1

Clinical example 2

Patient B., 60 years were treated at the cardiology Department of the MUSES "the 3rd hospital" with 13.10.2007 on 2.11.2007 years with a diagnosis of ischemic heart disease, acute Q-myocardial infarction anterior-septal area of the left ventricle from 13.10.2007 year. Hypertension Art. III, HSN 2 tbsp., FC 3, diabetes mellitus type 2, moderate, decompensation, risk 4. CHD diagnosed in 2006.

When hemorheological examination level POE amounted comprised 17.54%, II refers to the severity of the aggregation disorders. Other rheological parameters are as follows: average unit size - 7,12, the rate of aggregation - 3,43, blood viscosity at the shear rate of 200 s-1- 5.1 MPa·s, at a speed of 50-1- 6,2 MPa·s at shear rate 10s-1-10,1 MPa·s, the viscosity of the plasma - 1.8 MPa·s; hematocrit - 41,2%.

Clinical example # 3

Patient L., 67 years of age, was treated at the cardiology Department of the MUSES "the 3rd hospital" with 14.10.2006 on 12.11.2006 years with a diagnosis of coronary artery disease, recurrent Q-myocardial infarction anterior-septal area of the left ventricle from 14.10.2006 years, complicated by pulmonary edema. Hypertension Art. III, HSN 2A Art., FC 3, diabetes mellitus type 2, moderate, decompensation, risk 4. CHD diagnosed in 2002.

When hemorheological examination POE was -
3.97%, which refers to III severity aggregation disorders. Other rheological parameters are as follows: average unit size - 11,46, the rate of aggregation - 8,10, blood viscosity at the shear rate of 200 s-1at 5.6 MPa·s, at a speed of 50-1to 6.5 MPa·s at a shear rate of 10-1is 19.2 MPa·s, the viscosity of plasma is 2.7 MPa·s; hematocrit - 49,8%.

Thus, these examples demonstrate the feasibility of using the percentage of non-aggregated RBCs to assess the severity of the violation aggregation of erythrocytes.

A method of evaluating the severity of the violation aggregation of erythrocytes, including the definition of clinical and laboratory parameters, characterized in that in the blood plasma after 4 min after the beginning of the spontaneous aggregation of red blood cells to determine the number of free cells, and the number of cells in aggregates, then calculate the percentage of non-aggregated RBCs (POE) according to the formula POE=CSE·100/(sea+CTU), where CTU - free erythrocytes, sea - sum of all cells in the aggregates, and POE from 56 to 30% - I set the severity, from 30% to 4% - II the severity, less than 4% - III severity.

SUBSTANCE: thrombosis monitor comprises: a thrombosis chamber, at least in a part of which there is a thrombogenic material; an inlet tube connected to the thrombosis chamber through which blood flows into the thrombosis chamber; a blood supply container connected to the inlet tube; a feed pump for the container; a pressure sensor for measuring pressure applied to the container. A method of thrombosis monitoring consists in the fact that after introduction of an anticoagulant, blood is supplied from the container to the thrombosis chamber by pressing on a fluid placed on a blood layer and having density less than that of the blood layer; it is combined with anticoagulation blood processing or blood coagulation stimulation, and measurement of pressure applied to the container; the thrombogenic material is placed at least in a part of the thrombosis chamber.

EFFECT: group of inventions provides overall assessment of blood coagulation and platelet-cell thrombosis in a medium equivalent to blood flow for evaluation of efficacy of an antithrombotic drug.

SUBSTANCE: for determination of functional state of hemostasis system record of blood coagulation process is performed, current amplitude of blood resistance in first time moment is registered and second resistance of blood at multiple time moment from initial time value is measured. Two resistances and time moments are used to determine maximum blood resistance and time constant, by which blood resistance at the beginning and end of coagulation process is calculated. Obtained parameters are used to determine indices of beginning and end of blood coagulation process. Obtained indices are compared with of the same name indices of blood coagulation process in norm and in case of differently directed deviations disturbances of functional state of hemostasis system are diagnosed.

SUBSTANCE: method is based on a method of observing turbidimetric fibrin clot formation with optical transmission of an incubation medium recorded by ultraviolet radiation band 230 to 320 nm by means of UV-spectrophotometre as a fibrin-polymer detector.

EFFECT: invention enables higher accuracy and sensitivity of the method.

SUBSTANCE: for thrombin production measurement, a layer of said sample contacts with a fluorogenic substratum of thrombin where the thickness of said layer is 0.05 to 5 mm, while the surface area is 10 to 500 mm2. Further, the thrombin production environment in said sample is provided. It is followed by measuring the fluorescence emitted from the layer surface by a fluorescent group released by the fluorescent substratum as a result of an enzymatic action of produced thrombin on said fluorogenic substratum. Besides, the invention ensures a kit for measuring the thrombin activity in the sample.

SUBSTANCE: blood is examined. A hematocrit level (H), erythrocyte count (E), thrombocyte count (T) are determined. Said parametres are evaluated. In the event if they keep within the determined limits for the patients with acute coronary syndrome (ACS), then adenosine phosphate induced (ADP-induced) clotting time test samples are prepared. Citrated blood sample 0.4 ml is prepared of whole blood and divided on two samples 0.2 ml. Each of these samples is introduced in a measuring cell, recalcified at temperature 37°C for 2 minutes. Then a magnetic ball mixer is placed in each cell. The measurement is activated, and in three seconds the ADP solution 0.1 ml is introduced. After a clotting reaction, a process time duration is recorded separately for each sample. An arithmetical mean of the derived values is calculated (A). The derived values of each of said parameters are scored. Total score Σ=A+H+E+P shows the risk of recurrent thrombotic events. If Σ=4 points, the low risk is observed; the value Σ=5-6 points shows the medium risk, while Σ=7-10 points - the high risk.

EFFECT: method provides more objective risk evaluation of recurrent thrombotic events in the patients with ACS with its simplicity and low cost.

SUBSTANCE: blood sample is placed in capillary, in whose walls installed are electrodes connected to frequency generator and registering unit, blood electric conductivity is measured at the moment of passing through it of alternating current with frequency 200 Hz, electric coagulogram is registered and used to determine chronometric and amplitude characteristics: A - amplitude of functional curve decline, mV; N - time of functional curve decline to minimal value in minutes. If value of A/T index decreases or increases with respect to normal, conclusion about hemostatic disorders is made. If value of A/T index equals 3-5 - hemostasis state is evaluated as normal, if A/T value is lower than 3, hypocoagulation is determined, and if A/T value is higher than 5 - hypercoagulation.

EFFECT: application of the method makes it possible to obtain data about hemostasis system state in real time mode, without injuring form blood elements in investigated microvolumes of blood, thus making it possible to increase accuracy, self-descriptiveness and efficiency of hemostasis state evaluation and to carry out correction of performed therapy without delay.

SUBSTANCE: in patients with IHD before therapy with acetylsalisylic acid (ASA) ADP-induced and ASA-dependent platelet aggregation are examined and by their difference value of coefficient of aggregation inhibition (CAI) is calculated. CAI value <24% testifies to resistance to ASA, if CAI ≥24% - about sensitivity to ASA.

EFFECT: method ensures high prediction accuracy and makes it possible to prevent development of undesirable coronary events in IHD patients.

SUBSTANCE: invention refers to medicine, namely to haematology, and orthopaedics. The intravascular thrombocyte activity correction in children with scoliosis requires the prescription for at least 6 months of a therapeutic complex which involves daily therapeutic physical training, two courses of massage and daily swimming for at least 20 minutes a day.

EFFECT: method allows to normalise intravascular thrombocyte activity in children aged 8-12 with scoliosis, to improve tissue microcirculation considerably, to optimise growth and development of skeleton and internals.

SUBSTANCE: invention belongs to medicine, notably to haematology and orthopaedics. For thromboplastin synthesis correction in children 8-12 years old with scoliosis is prescribed complex of treatment, including daily remedial gymnastics, two courses of massage and daily swimming for at least 20 minutes a day, during 6 months.

EFFECT: method enables thromboplastin synthesis normalisation in children with scoliosis, significantly sanitate children with scoliosis due to improvement of tissues' microcirculation.

SUBSTANCE: the suggested studying should be carried out on the glass simultaneously with several inductors by applying minimal inter-taking antilogarithms concentrations of aggregation inductors which correspond at double combination of inductors: ADP 5.0 x 10-8 M, adrenaline 3.0 x 10-9, collagen - dissolving the main suspension 1:8, thrombin 0.075 U/ml; at triple combination of inductors: ADP 10-9 M, adrenaline 10-9, collagen - dissolving the main suspension 1:9, thrombin 0.060 U/ml. The development of aggregation means thrombocytic activation in patients with arterial hypertension at metabolic syndrome. The method enables to evaluate the changes of thrombocytic functional state with combination of inductors more probably present in area of vascular lesion by applying minimal necessary concentrations that develops real conditions at hemostatic initiation in human vessels.

SUBSTANCE: one should evaluate the time for clotting of plasma under testing in phospholipid-dependent test, moreover, one should apply high- and low-sensitive thromboplastin reagents to lupus anticoagulant to calculate the ratio of indices of prothrombin time prolongation and at its value being either equal to or above 1.1 one should diagnose APS.

SUBSTANCE: one should study blood components to detect anticoagulant-fibrinolytic activity. Moreover, patient's blood should be sampled: in whole blood one should detect the presence of affected erythrocytes and evaluate the quantity of thrombocytes, in plasma it is necessary to study the activity of antithrombin III, XIIa-dependent fibrinolysis, the content of soluble fibrin-monomeric complexes, in blood serum of the sample taken one should detect the concentration of urea, creatinine, sodium, albumin, total cholesterol and the activity of aspartate aminotransferase, moreover, one should calculate integral value of renal-hepatic deficiency, to put corresponding point for the degree of parameters under testing, then one should calculate integral value of disseminated intravascular clotting (IVDIC) and at its value being 6.3 U and more DIC-syndrome should be diagnosed, moreover, at IVDIC value ranged 6.3-10.1 U it is possible to diagnose latent DIC-syndrome, at 10.2-14.6 - subacute DIC-syndrome and at 14.7 and higher - acute DIC-syndrome should be concluded.

SUBSTANCE: the present innovation deals with predicting disadaptive processes in women in dynamics of menstrual cycle. During menstrual cycle beginning since the 1st d to the 21st d one should detect the dynamics for alteration in coefficient of activity of syntoxic adaptation programs (CASAP), calculated by the following formula:

where CST - concentration of blood serotonin, AAT-III - activity of antithrombin III, Aaoa - total antioxidizing activity of plasma, CCD8+ - concentration of T-suppressors, Cad - concentration of blood adrenalin, Cα2MG - concentration of α2-macroglobulin, CMDA - concentration of malonic dialdehyde, CCD4+ - concentration of T-helpers. Moreover, normally CASAP value alters two-fold against the first day of the cycle - since 0.70 up to 1.40 on the 21st d of the cycle, at no alterations in CASAP value one should diagnose female disadaptive alterations leading to failed pregnancy. The innovation enables to perform diagnostics of disadaptive processes in women in dynamics of menstrual cycle followed by prognostic conclusion upon future pregnancy.

SUBSTANCE: method involves determining coagulating blood viscosity values like reaction period r, thrombin constant K, maximum amplitude MA, time T for forming fibrin-thrombocytic blood clot, spontaneous blood platelets aggregation intensity Ar, retraction and spontaneous clot lysis total FA. The r being within 5-7 min, Ar from -2 to -6 relative units, K being within 4-6 min, MA within 500-700 relative units, T within 40-60 min and FA equal to 10-20%, low inflammatory process activity is considered to be the case. The r being less than 5 min, Ar equal to -8 to -12 relative units, T less than 40 min and FA less than 10% with no changes in K and MA being observed, inflammatory process activity in chronic glomerulonephritis case is considered to be of high severity degree.

SUBSTANCE: one should study both activation and aggregation of thrombocytes in blood of carotid artery, at the quantity of thrombocytic active forms being above 70% and the number of aggregated thrombocytes being above 9.0% one should predict the development of cerebral ischemic lesion along with stable focal neurological symptomatology, and at the quantity of thrombocytic active forms being below 30% and the number of aggregated thrombocytes being below 8.0% it is possible to predict positive dynamics in the course of the disease mentioned without developing cerebral ischemic lesion.

SUBSTANCE: one should study the level of von Willebrand's factor in patient's carotid artery blood. At its content being below 105% one should predict the development of repeated AICH. The innovation improved information value of testing due to possibility to obtain reliable prediction in latent period, as well.